LOCUS       sequence-426367-        4206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4206)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 4206)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221-SLC22A4_STOP
FEATURES             Location/Qualifiers
     source          1..4206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1763..1862)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1879..1898)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1880..1898)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1903..1919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1903..1919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2021..2040)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2032..2841
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2099..2118)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2988..3576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3477..3496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3730..3747
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3906..3933)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4025..4111)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4160..4182
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4174..4191
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4175..4191
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgcg ggactacgat
      121 gaagtgatcg cctttctggg cgagtggggc cctttccaga gactgatctt ctttctgctg
      181 tctgccagca tcatcccaaa cggctttaat ggcatgtctg tagtatttct tgcaggaacc
      241 cctgagcaca ggtgcagagt gccagacgca gcaaacctga gctccgcctg gaggaacaat
      301 agcgtgccac tgaggctgcg cgatggcaga gaggtgcccc actcctgttc tcggtataga
      361 ctggccacaa tcgccaattt ttccgccctg ggactggagc caggaaggga cgtggatctg
      421 ggacagctgg agcaggagtc ttgcctggac ggctgggagt tctctcagga cgtgtacctg
      481 agcaccgtgg tgacagagtg gaacctggtg tgcgaggaca attggaaggt gcccctgacc
      541 acatctctgt tctttgtggg cgtgctgctg ggctcttttg tgagcggcca gctgagcgat
      601 cggtttggca gaaagaacgt gctgttcgcc accatggccg tgcagacagg cttcagcttt
      661 ctgcagatct tcagcatctc ctgggagatg ttcaccgtgc tgtttgtgat cgtgggcatg
      721 ggccagatct ccaattatgt ggtggccttt atcctgggca cagagatcct gggcaagtcc
      781 gtgaggatca tcttctctac cctgggcgtg tgcacattct ttgccgtggg ctacatgctg
      841 ctgcctctgt tcgcctactt catcagggac tggaggatgc tgctgctggc cctgaccgtg
      901 ccaggcgtgc tgtgcgtgcc actgtggtgg tttatccccg agagccctag gtggctgatc
      961 tcccagcgga gattccgcga ggccgaggat atcatccaga aggccgccaa gatgaacaat
     1021 atcgccgtgc ctgccgtgat cttcgactcc gtggaggagc tgaacccact gaagcagcag
     1081 aaggccttca tcctggatct gtttaggacc cgcaatatcg ccatcatgac aatcatgtct
     1141 ctgctgctgt ggatgctgac ctctgtgggc tactttgccc tgagcctgga cgccccaaac
     1201 ctgcacggcg atgcctacct gaattgcttc ctgagcgccc tgatcgagat ccccgcctat
     1261 atcaccgcct ggctgctgct gaggacactg cctaggcgct acatcatcgc agccgtgctg
     1321 ttttggggag gaggcgtgct gctgttcatc cagctggtgc ccgtggacta ctattttctg
     1381 agcatcggcc tggtcatgct gggcaagttc ggcatcacat ccgccttttc tatgctgtac
     1441 gtgttcaccg ccgagctgta tcctacactg gtgaggaaca tggcagtggg agtgaccagc
     1501 acagcatcca gggtgggatc catcatcgca ccatacttcg tgtatctggg cgcctacaat
     1561 cgcatgctgc cctatatcgt gatgggcagc ctgaccgtgc tgatcggcat cctgacactg
     1621 ttctttcccg agtccctggg catgaccctg cctgagacac tggagcagat gcagaaggtg
     1681 aagtggtttc ggtccggcaa gaagaccaga gattctatgg agacagagga gaaccccaag
     1741 gtgctgatca ccgccttcta agacccagct ttcttgtaca aagttggcat tataagaaag
     1801 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     1861 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     1921 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1981 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     2041 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     2101 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     2161 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     2221 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     2281 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     2341 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     2401 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     2461 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     2521 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     2581 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2641 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2701 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2761 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2821 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     2881 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     2941 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3001 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3061 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3121 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3181 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3241 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3301 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3361 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3421 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3481 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3541 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3601 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3661 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3721 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3781 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3841 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     3901 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     3961 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     4021 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     4081 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
     4141 acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc
     4201 gggccc
//
